Anti-TFEB antibody [BLR070G]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-TFEB antibody [BLR070G] (AB264421)

Flow cytometry overlay histogram of 4% formaldehyde fixed Daudi cells permeabilized with 90% methanol labeling human TFEB with ab264421 at 1 µl per 1 x 10^6 cells (shaded) compared with a Isotype control (unshaded). Secondary antibody details : Goat Anti-Rabbit IgG H&L (DyLight® 650) (ab96886).

• IP

Supplier Data

Immunoprecipitation - Anti-TFEB antibody [BLR070G] (AB264421)

TFEB was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) with ab264421 at 20 μl/mg lysate. Western blot was performed from the immunoprecipitates using ab264421 at 1/1000 dilution.

Lane 1 : ab264421 IP in HeLa whole cell lysate.
Lane 2 : Control IgG IP in HeLa whole cell lysate.

Detection : Chemiluminescence with an exposure time of 75 seconds.

Lysates prepared using NETN lysis buffer.

All lanes:

Immunoprecipitation - Anti-TFEB antibody [BLR070G] (ab264421)

Predicted band size: 53 kDa

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• WB

Lab

Western blot - Anti-TFEB antibody [BLR070G] (AB264421)

False colour image of Western blot : Anti-TFEB antibody [BLR070G] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab264421 was shown to bind specifically to TFEB. A band was observed at 60 kDa in wild-type HepG2 cell lysates with no signal observed at this size in TFEB knockout cell line ab277871 (knockout cell lysate ab284220). To generate this image, wild-type and TFEB knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-TFEB antibody [BLR070G] (ab264421) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

TFEB knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human TFEB knockout Hep G2 cell line (<a href='/en-us/products/cell-lines/human-tfeb-knockout-hep-g2-cell-line-ab277871'>ab277871</a>)

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 53 kDa

Observed band size: 60 kDa

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• WB

Supplier Data

Western blot - Anti-TFEB antibody [BLR070G] (AB264421)

Lysates prepared using NETN lysis buffer.

All lanes:

Western blot - Anti-TFEB antibody [BLR070G] (ab264421) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

A549 whole cell lysate at 50 µg

Lane 3:

HepG2 whole cell lysate at 50 µg

Lane 4:

Jurkat whole cell lysate at 50 µg

Lane 5:

RKO whole cell lysate at 50 µg

Lane 6:

K562 whole cell lysate at 50 µg

Lane 7:

LNcaP whole cell lysate at 50 µg

Lane 8:

MCF7 whole cell lysate at 50 µg

Lane 9:

TCMK-1 whole cell lysate at 50 µg

Lane 10:

NIH/3T3 whole cell lysate at 50 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG

Predicted band size: 53 kDa

true

Exposure time: 3min